Objective:To screen for the pathogenesis-related genes of osteosarcoma and to assess their roles for the de- velopment of osteosareoma.Methods:Total RNA was extracted from 3 ATCC osteosarcoma cell lines and an osteoblastic cell line and was used to synthesize biotinylated cRNAs;the latter were hybridized to Affymetrix~(?)GeneChip~(?)U133A ar- rays and a gene with more than 2 folds of change was selected.Ten of the differentially expressed genes were chosen and the primers were designed and the synthesized.Then SYBR~(?)Green real-time PCR(RT-PCR)method was used to detect the expression of the 10 genes in 9 fresh osteosarcoma specimens.ABI Prism 7 000 system was used to analyze the differ- ent expression between osteosarcoma cell line and osteoblastic cell line.Results:We identified 58 up-regulated and 142 down-regulated genes in the 3 osteosareoma cell lines.Many of the genes were firstly reported to be related to the patho- genesis of osteosarcoma.These differentially expressed genes were mainly involved in energy and material metabolism,on- cogene,signal transduction gene,transcription- related genes,cell cycle-related genes,cell apoptosis-related gene,im- mune response gene,tumor suppressor genes,etc.The array results of 10 randomly selected genes were further verified by the RT-PCR in 9 fresh osteosarcoma specimens.Conclusion:Many genes are involved in the pathogenesis of osteosarcoma. Gene microarray can help to discover the genes related to the pathogenesis of osteosarcoma,which may lay a foundation for studying the molecular mechanism of osteosarcom.

OBJECTIVETo observe the effect of umbilical sticking therapy (UST) with Qitou Xiaugu Plaster (QXP) on hemodynamics of portal system in patients with liver cirrhosis.

METHODSOne hundred and twenty patients of liver cirrhosis with portal hypertension were assigned to two groups. On the basis of conventional therapy, UST was applied in the 66 patients in treated group, which was exchanged once every 3 days with an interval of 1-day rest. The 54 patients in the control group were orally administered with propanolol. The therapeutic course for both groups was 1 month. Before and after treatment, the hemodynamic changes in portal or splenic veins were observed by color Doppler ultrasonograph, and the changes of liver function, blood coagulation and patients' subjective symptoms were observed as well.

RESULTSAfter treatment, portal vein diameter and splenic vein diameter significantly decreased (P < 0.05, portal venous flow velocity and splenic venous flow velocity apparently increased (P < 0.05), and portal venous flow apparently decreased in both groups (P < 0.05), while no significant change was found in the splenic venous flow (P > 0.05). The liver function and blood coagulation indexes in both groups were improved. The improvement of clinical symptoms in the treated group was superior to that in the control group.

CONCLUSIONUST with QXP could decrease the portal vein pressure in a short time, with the therapeutic effect comparable to propanolol, and with no adverse reaction.

Adult ; Aged ; Drugs, Chinese Herbal ; administration & dosage ; Female ; Hemodynamics ; drug effects ; Humans ; Hypertension, Portal ; drug therapy ; physiopathology ; Liver Cirrhosis ; drug therapy ; physiopathology ; Male ; Middle Aged ; Portal Pressure ; drug effects ; Portal Vein ; drug effects ; physiopathology ; Splenic Vein ; drug effects ; physiopathology ; Umbilicus ; blood supply ; Young Adult

OBJECTIVE: To observe the expression pattern of caspase-3 and HCLS1-associated protein X-1 (HAX-1) at different time after cerebral contusion in rat, and explore the new method for estimating the injury interval. METHODS: The cerebral contusion model was established using adult SD male rats. Then the rats were randomly allocated into 8 groups: 2 h, 6 h, 12 h, 1 d, 3 d, and 7 d after cerebral contusion, sham-operation and normal control. Expression of caspase-3 and HAX-1 protein after cerebral contusion in rat was detected by Western blotting. Laser scanning confocal microscope was used to observe the number of HAX-1 positive cells and TUNEL-stained cells after cerebral contusion. RESULTS: The expression of caspase-3 increased parallelly with the time after cerebral contusion and reached the peak value on 3 d. The expression of caspase-3 decreased gradually and still maintained a high level expression on 7 d (P < 0.05). The expression of HAX-1 positive cell went up after injury, and reached the peak value at 6 h (P < 0.05), then turned down gradually after 12 h and went out of detection after 3 d. The number of TUNEL-stained cells increased obviously at 2 h and reached the peak value on 3 d. The number of TUNEL-stained apoptotic cells decreased gradually and still maintained a high level expression on 7 d (P < 0.05). CONCLUSION The expression of caspase-3 and HAX-1 after cerebral contusion has time sequential regularity, which may provide new evidence for forensic diagnosis of cerebral contusion interval.
Animals ; Blotting, Western ; Brain Injuries/pathology* ; Carrier Proteins/metabolism* ; Caspase 3/metabolism* ; Cerebellum/pathology* ; In Situ Nick-End Labeling ; Intracellular Signaling Peptides and Proteins ; Male ; Rats ; Rats, Sprague-Dawley

Objective:To investigate the effects of estradiol on ~(60)Co?-ray induced apoptosis of bone marrow hematopoietic cells of mice,and to discuss the related anti-irradiation mechanism.Methods:KM mice were randomly divided into 3 groups(15 mice/each group):control group(without radiation),pure radiation group and estradiol+radiation group(ER group).The pure radiation group was irradiated by 4.0 Gy?-ray at a dose rate of 1.15Gy/min;the ER group was administered with 0.1 mg estradiol(IM)at 10 days before 4.0 Gy?-ray radiation;and the control group received no special treatment.The apoptotic DNA segments of bone marrow hematopoietic cells were analyzed by DNA agarose gel electrophoresis;flow cytometry was used to examine the apoptosis rate of cells and expression of Fas and Bcl-2 at 4 h,8 h,and 12 h after irradiation.Results:Eight hours after radiation,the apoptotic DNA segments were obviously increased and apoptotic DNA ladder appeared,which was not seen in the other 2 groups.The apoptosis rate of bone marrow hematopoietic cells in ER group was significantly lower than that in the pure radiation group at 4,8,and 12 h after irradiation(P

Animals ; Humans ; Oxazines ; metabolism ; Phenylpropionates ; metabolism ; Receptors, Cytoplasmic and Nuclear ; agonists ; antagonists & inhibitors ; Thiazolidinediones ; metabolism ; Transcription Factors ; agonists ; antagonists & inhibitors

Cell Culture Techniques ; methods ; Fixatives ; Humans ; Immunohistochemistry ; methods ; Tissue Fixation ; methods

Objective To investigate the clinical feature,treatment,and prognosis of hospitalized patients with jaw osteoradionecrosis. Methods A total of 93 cases with jaw osteoradionecrosis treated between 2000 and 2010 was reviewed.Of the 93 cases,79 cases were with mandible lesions,13 cases with maxillary lesions,and 1 case with both mandible and maxillary lesions.Sixty-six cases received one course of radiotherapy,with the radiation doses of 34 -90 Gy (mean 64.6 Gy).Twenty-two cases experienced tooth extraction or other operative procedures before exhibition of the clinical symptoms for osteoradionecrosis.The interval time between radiotherapy and the onset of osteoradioneerosis varied from 2 weeks to 32 years (mean 54 months).Results Of 93 cases,56 patients underwent radical resection of the pathologic bone and reconstruction with free tissue flaps,in whom 7 cases received the second surgery due to microvascular thrombosis in flap vessels,and flaps were survival by new vascular anastomosis in 3 cases,the failed flaps were removed and replaced successfully by non vascularized bone grafts in 2 cases,and the failed flaps removed and the defects were repaired with adjacent skin in other 2 cases.In the 56 cases,only one case was with disease recurrence and 53 cases with significant improvement in chewing and swallowing functions.Only 2 of 93 cases underwent resection of the pathologic bone and reconstruction with titanium plates,and thereafter they encountered titanium exposure.Scaling of osteoradionecrosis lesions was applied to 20 of 93 patients and 9 cases of them were with disease recurrence.Fifteen cases had resection of the effected mandible without reconstruction.Disease relapse was encountered in 2 of them,others had poor chewing and swallowing.Conclusions The mandible is more susceptible to osteoradionecrosis than maxilla.Radical resection with reconstruction by free tissue flap is recommended for the treatment of jaw osteoradionecrosis,and scaling and reconstruction only with titanium plate should be avoided because of high risks of titanium exposure and disease relapse.

Objective To study the protective effect of Irisin in doxorubicin(Dox)induced-Cardiomyopathy and its possible mechanism.Methods AC 16 cells were used to construct Dox injury model and divided into control group(AC 16 cells were cultured with complete medium),Irisin group(AC16 cells were treated with 10 ng·L-1 Irisin for 24 h),Dox group(AC 16 cells were treated with 4 μmol·L-1 Dox for 24 h),Dox+Irisin group(AC 16 cells were pretreated with 10 ng·L-1 Irisin for 2 h,and then treated with 4 pmol·L-1 Dox for 24 h).Cell counting kit-8(CCK-8),terminal deoxynucleotidyl transferase-mediated nick end labeling(TUNEL)and lactate dehydrogenase(LDH)were used to detect the proliferation,apoptosis and mortality of AC 16 cells.Western blot was used to detect the expression levels of nuclear factor-κB(NF-κB)signaling pathway and apoptotic factors B-cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax)and caspase-9 protein.Mito-Tracker Red CMXRos probe was used to detect mitochondrial membrane potential.Results In the contrl group,Irisin group,Dox group,Dox+Irisin group,the rate of apoptosis were(0.97±0.09)％,0,(42.80±6.70)％,(11.74±1.79)％;the expression of Bax protein were 0.85±0.01,0.36±0.02,1.15±0.07,0.37±0.11;the expression of caspase-9 protein were 0.52±0.02,0.59±0.03,1.11±0.02,0.67±0.08;the expression of Bcl-2 protein were 1.01±0.04,1.05±0.25,0.43±0.02 and 0.99±0.30;the probability of mitochondrial damage were(0.02±0.01)％,(0.5±0.15)％,(38.6±2.39)％,(1.58±0.54)％.The difference of the above indexes between the contrl group and the Dox group were statistically significant(all P＜0.05);the difference between Dox group and Dox+Irisin group were statisically significant(all P＜0.05).Conclusion Irisin could reduce the expression level of Bax,caspase-9,p-NF-κB,and p-mTOR caused by Dox,increase the expression level of Bcl-2,ameliorate the myocardial damage caused by Dox,and reduce cardiotoxicity.

Objective To explore the causes of twisted or tailed DNA bands in agarose gels after electro-phoresis .Methods Prestained and poststained methods were employed to exam 3 kinds of DNA marker bands in agarose gel with GeneGreen and Ethidium Bromide ,respectively .Results Three kinds of DNA marker bands in agarose gel with 1:10000 and 1:5000 concentration of GeneGreen showed obvious distortions and trailing phenomena compared with the same concentration of Ethidium Bromide w hen the prestained method was used for electrophoresis ,which were improved when the poststained method was used in agarose gel with GeneGreen and Ethidium Bromide ,respectively .Conclusion Nucleic acid dye GeneGreen can affect the quali-ty of DNA bands in agarose gel electrophoresis .When the quality of DNA itself ,agarose gel quality ,electro-phoretic fluid quality and voltage are excluded ,the quality of nucleic acid dye should be taken into considera-tion if the bands twisting or tailing occurs when the DNA nucleic acid electrophoresis is carried out by pres-tained method .The quality of DNA electrophoresis bands can be improved by using poststained method or re-placing nucleic acid dye .

OBJECTIVE: To investigate the expression of caspase-3 and iNOS in different intervals and to provide evidence for estimation of injury intervals after brain contusion in human. METHODS: Thirty cases died of serious brain injury were included into the injury groups and 5 cases died of non-brain injury were served as control group. To analyze the changes of caspase-3 and iNOS expression in brain samples at different intervals (2h, 4-8h, 10-14h, 1-2d, 3-5d, 8-11d) by immunohistochemistry and auto-image analysis system. RESULTS: The level of caspase-3 expression started to increase in 2 hours after brain contusion compared to the control group (P<0.05). The level of caspase-3 expression continued to increase in 1-2 days and maintained high level in 3-5 days compared to the control group (P<0.05), then decreased gradually. There was no statistically significant difference between the expression level of iNOS in 2 hours with the control group (P>0.05). But the expression level of iNOS began to increase in 4-8 hours after brain contusion and reached its maximum in 1-2 days, then decreased. Weak expression of iNOS still could be detected in 8-11 days. CONCLUSION The expression of caspase-3 and iNOS can be used as effective evidence for human brain contusion interval.
Adult ; Astrocytes/metabolism* ; Brain/pathology* ; Brain Injuries/pathology* ; Caspase 3/metabolism* ; Female ; Forensic Pathology ; Humans ; Immunohistochemistry ; Male ; Middle Aged ; Neurons/metabolism* ; Nitric Oxide Synthase Type II/metabolism* ; RNA, Messenger/metabolism* ; Staining and Labeling ; Time Factors ; Young Adult